CAJ | 학술논문

泛素结合酶（UbcE2）是蛋白泛素化过程中所必需的酶，在泛素转移和底物的特异性识别方面发挥着重要的作用。本实验利用 RACE 技术克隆获得了全长为993 bp 的松江鲈泛素结合酶 E2-D2基因 cDNA 序列（命名为 TfUbcE2-D2），其开放阅读框为444 bp，编码147个氨基酸。通过 SMART 预测得知，TfUbcE2-D2含有一个 UBCc 结构域。同源比对结果表明该基因与其他物种的同源性为92．31％。实时荧光定量 PCR 显示，TfUbcE2-D2广泛表达于松江鲈各组织，在肾脏中的相对表达量最高，其次为鳃。鳗弧菌（Vibrio anguillarum ）刺激后，TfUbcE2-D2在血液、脾脏、肝脏和鳃中表达均上调，其中，脾脏和鳃中的表达量上调约40倍。由以上实验结果推测，TfUbcE2-D2可能参与松江鲈的先天免疫防御。
범소결합매（UbcE2）시단백범소화과정중소필수적매，재범소전이화저물적특이성식별방면발휘착중요적작용。본실험이용 RACE 기술극륭획득료전장위993 bp 적송강로범소결합매 E2-D2기인 cDNA 서렬（명명위 TfUbcE2-D2），기개방열독광위444 bp，편마147개안기산。통과 SMART 예측득지，TfUbcE2-D2함유일개 UBCc 결구역。동원비대결과표명해기인여기타물충적동원성위92．31％。실시형광정량 PCR 현시，TfUbcE2-D2엄범표체우송강로각조직，재신장중적상대표체량최고，기차위새。만호균（Vibrio anguillarum ）자격후，TfUbcE2-D2재혈액、비장、간장화새중표체균상조，기중，비장화새중적표체량상조약40배。유이상실험결과추측，TfUbcE2-D2가능삼여송강로적선천면역방어。
Ubiquitin-conjugating enzyme is the second enzyme in the process of protein ubiquitination and plays an important role in ubiquitin transferring and substrate specific recognition.In this study,we cloned the full-length cDNA from the Roughskin sculpin,Trachidermus fasciatus by RACE,and named as TfUbcE2-D2.The cDNA of TfUbcE2-D2 is 993 bp,which contained an open reading frame (ORF)of 444 bp encoding a polypeptide of 147 a-mino acids.The deduced amino acid sequence had a domain called Ubcc,which was extremely conservative.The mRNA expression of TfUbcE2-D2 in healthy andVibrio anguillarum challenge was detected by quantitative real-time PCR,results showed that TfUbcE2-D2 was expressed in all the detected tissues,mainly expressed in kidney and gill.The up-regulated of TfUbcE2-D2 post challenge with Vibrio anguillarum in blood,spleen,liver and gill indicated that it might be involved in the innate response of Roughskin sculpin.